ClickTech EdU Cell Proliferation Kit 594 (100 Assays) for IM
EdU Cell Proliferation Kit for Imaging
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All EdU based proliferation kits on the market are patented by our company since 2007. Using EdU shows several advantages over other cell proliferation assays (BrdU or 3H-thymidine) making it a superior evaluation tool for monitoring genotoxicity, evaluation of anticancer drugs or cell cycle analysis. Measured are only the cells in vitro and/or in vivo that are actively dividing. Healthy cells that are not dividing are not detected and can thus give no false positives. The assay can be used for single cell proliferation analysis as well as cell division in animals. After incorporation of EdU, cell proliferation can be detected in different organs. The amount of EdU used in the assays shows no toxicity in cells as well as in animals thus does not influence the results of experiments.
The kits are designed for fluorescence microscopy. They contain special reagents and a dedicated protocol for read-outs using fluorescence microscopy on either cells in suspension or attached cells.
The kits are available for the following wavelenghts: 488 nm; 555 nm; 594 nm; 647 nm
LITERATURE
Anticancer drug and ionizing radiation-induced DNA damage differently influences transcription activity and DDR-related stress responses of an endothelial monolayer, V. Ziegler et al., 2020, Molecular Cell Research, Vol. 1867, p. 118678.
https://doi.org/10.1016/j.bbamcr.2020.118678
Dendrimer-Based Signal Amplification of Click-Labelled DNA in Situ, N. Raddaoui et al. 2017, ChemBioChem, Vol. 18, p. 1716-1720.
https://doi.org/10.1002/cbic.201700209
High-copy sequences reveal distinct evolution of the rye B chromosome, S. Klemme et al., 2013, New Phytologist, Vol. 199, p. 550-558.
https://doi.org/10.1111/nph.12289
Secretome analysis of patient-derived GBM tumor spheres identifies midkine as a potent therapeutic target, S. Han et al., 2019, Experimental & Molecular Medicine, Vol. 51(12), p. 1-11.
https://doi.org/10.1038/s12276-019-0351-y
Small extracellular vesicles from hypoxic mesenchymal stem cells alleviate intervertebral disc degeneration by delivering miR-17-5p, Z. M. Zhou et al., 2022, Acta Biomaterialia, Vol. 140, p. 641-658.
https://doi.org/10.1016/j.actbio.2021.11.044
BCL-6 promotes the methylation of miR-34a by recruiting EZH2 and upregulating CTRP9 to protect ischemic myocardial injury, J. M. Lin et al., 2021, BioFactors, Vol. 47(3), p. 386-402.
FAQ
- What type of cells can incorporate EdU?
The EdU cell proliferation assay has been applied to many different cell types and organisms from prokaryotic to eukaryotic. Cell lines such as E. coli, HeLa, HEK, MOLM are arguably among the most routine applications, but also animals, like mouse, rat, the nematode C. elegans, crickets (Gryllus bimaculatus), chicken (Gallus domesticus) and zebra fish (Danio rerio) or even plants (e.g. Arabidopsis thaliana) can be applied.
Cells that possess a pyrimidine pathway that can phosphorylate EdU to the corresponding triphosphate, which is then accepted by the host DNA polymerase for incorporation into DNA during replication. - Can I perform EdU cell proliferation detection on living cells?
EdU is incorporated into living cells, but the detection reaction must be performed on fixed and permeabilized samples.
- How does EdU labeling compare to BrdU or the 3H-thymidine incorporation assay?
All three methods enable to determine cell proliferation directly by incorporation of a metabolite analogue and subsequent detection. The 3H-thymidine incorporation assay is very sensitive, but the radioactive compound requires specialized equipment and dedicated lab space for handling. EdU and BrdU assays are non-radioactive alternatives with decreased risk for health and environment. Compared to the BrdU incorporation assay the EdU assay is more sensitive, requires less handling time and needs no harsh DNA denaturing conditions for detection. Therefore, the EdU cell proliferation is also compatible with multiplexing.
- Can I combine DAPI staining and EdU detection?
Yes, this is feasible. Please note that DAPI staining should be done after the click detection step. Alternatively, SYBR Green DNA staining can be used. But, please note that SYBR green should not be used with dyes of 488 nm wavelengths.
- When can I safely interrupt the experiment?
It is possible to safely interrupt the protocol after the fixation step. Thereto, remove the fixation solution and wash as suggested by the user manual, then the cells can be stored in buffer at 4° C. Alternatively, the experiment can also be safely interrupted after permeabilization, again as described above.
Please note: It is important to proceed with the experiment if the click cocktail for the detection of the EdU has been prepared already. - Is antibody staining compatible with the EdU?
Antibody staining is compatible with EdU cell proliferation detection when antibody detection is done after the click detection step. Please be aware of the dye used for EdU detection. Check also the user manual for more information.
- How to determine the EdU incubation time?
The EdU incubation time depends on the cell type or organism, the applied EdU concentration and the experimental design. For a start it is advisable to refer to a literature protocol (which is close to your experimental setup) and to test the conditions with a low number of samples. As a general guideline we recommend to use a maximum of 10 µM final EdU in the cell culture medium for incubations. For longer incubation (> 1 day) the concentration should be decreased to 1-5 µM.
- What type of cells can incorporate EdU?
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- Shelf Life
12 months unopened after receipt
- Storage Conditions
2-8 °C
- Physical State
kit system made of different components
- CAS Number
n.a.
- Excitation (max)
584 nm
- Emission (max)
603 nm
- Ɛ (max)
110.000 cm-1M-1
- Preparation/Handling
please see user manual of the kit
- Shelf Life